A novel anti-p21Ras scFv antibody reacting specifically with human tumour cell lines and primary tumour tissues

BMC Cancer
Ju-Lun YangQiang Feng

Abstract

The ras genes play an important role in the development and progression of human tumours. Neutralizing Ras proteins in the cytoplasm could be an effective approach to blocking ras signalling. In this study, we prepared anti-p21Ras single chain fragment variable antibody (scFv) and investigated its immunoreactivity with human tumours. The coding sequences of H-ras, K-ras, and N-ras were separately ligated into the vector pET-28a(+). Then, recombinant expressing plasmids were induced by IPTG for p21Ras expression in E. coli. Hybridoma cell lines producing anti-p21Ras monoclonal antibodies were isolated using wildtype p21Ras proteins as immunogens. Anti-p21Ras scFv antibody was prepared from the hybridoma by the phage scFv display method. The immunoreactivity of the anti-p21Ras monoclonal antibody and the scFv antibody was identified by ELISA and immunocytochemistry. We prokaryotically expressed wildtype H-p21Ras, K-p21Ras and N-p21Ras and generated the hybridoma cell line KGH-R1, producing anti-p21Ras monoclonal antibodies. It was demonstrated that KGH-R1 monoclonal antibody could recognize wildtype and mutated H-p21Ras, K-p21Ras and N-p21Ras in human tumour cell lines. In all 14 types of primary human cancer tissues tested, the ...Continue Reading

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Citations

Aug 30, 2017·SLAS Discovery·Behzad JafariSiavoush Dastmalchi
Aug 18, 2020·Journal of Biomolecular NMR·Christopher B MarshallMitsuhiko Ikura
Mar 6, 2017·International Journal of Molecular Medicine·Shuang BaiJu-Lun Yang
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Mar 1, 2020·Drug Discovery Today·Yunfeng LinLei Zhang

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Datasets Mentioned

BETA
M54968
BC005219

Methods Mentioned

BETA
ELISA
phage display
PCR

Software Mentioned

IgBLAST
ARRIVE
MODEL
SWISS

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